Enhancement of Escherichia coli cellulolytic activity by co-production of beta-glucosidase and endoglucanase enzymes

Cellulase is a group of enzymes (endoglucanase, exoglucanase and beta-glucosidase) required for cellulosic feedstock hydrolysis during bioethanol production. The use of recombinant cellulase is a strategy to reduce the enzyme cost. In this context, the present work describes the construction of a cellulase expression vector (pEglABglA), which allowed constitutive co-expression of endoglucanase A (EglA) from an endophytic Bacillus pumilus and the hyperthermophilic \u3b2-glucosidase A (BglA) from Fervidobacterium sp. in Escherichia coli . When compared to the non-modified strain DH5\u3b1, the recombinant Escherichia coli DH5\u3b1 (pEglABglA) reduced fivefold the viscosity of the carboxymethylcellulose medium (CMC-M). Also, it presented almost 30-fold increase in reducing sugar released from CMC-M, enabling the recombinant strain to grow using CMC as the sole carbon and energy source. When cultivated in rich media, specific growth rates of recombinant E. coli strains BL21, JM101 and Top10 were higher than those of DH5\u3b1 and DH10B strains. The constructed plasmid (pEglABglA) can be used as backbone for further cellulase gene addition, which may enhance even more E. coli cellulolytic capacity and growth rate

Relationship between power condition, agility, and speed performance among young roller hockey elite players

Purpose. The study was intended to describe the correlations between speed performance (11 m, 22 m, and 33 m) and agility test in skates with determinants of muscular power (squat jump, countermovement jump) in young Portuguese roller hockey athletes involved in a regional selection. Methods. A cross-sectional study was conducted among 10 male roller hockey players with mean (? SD) age of 14.20 (0.57) years, involved in the Portuguese national competition of under-15, making part of the final Oporto district selection of players to participate in inter-regional selections competition. Their mean (? SD) weight, height, body mass index, and sum of the skinfolds were 58.62 (8.78) kg, 165.72 (8.45) cm, 21.26 (1.52) kg/m2, 51.80 (14.91) mm, respectively. Furthermore, it strength was measured with squat jump and countermovement jump; sprinting time at 11 m, 22 m, and 33 m was determined, as well as time in an agility t-test, conducted in roller skating. Spearman correlation coefficients were calculated to test the associations. Results. Significant inverse correlations between vertical jumps, a practical method of power training monitoring, and linear velocity in skating (countermovement jump vs. speed, ?0.78) were found. Despite the increasing complexity of the agility test, a moderate inverse correlation with strength was observed, too (?0.48). Conclusions. Lower limbs explosive strength turned out a strong predictor of skating linear speed and agility among young roller hockey players, providing a simple evaluation tool of important determinants of performance.D915-7373-ED16 | Cesar LeaoN/

GROWTH, NUTRIENT AND TOTAL PHENOLS ACCUMULATION IN AUSTRALIAN RED CEDAR SEEDLINGS ( Toona ciliata ) INOCULATED WITH MYCORRHIZAL FUNGI

O presente estudo teve por objetivo avaliar a produ\ue7\ue3o de mudas de cedro-australiano quanto \ue0 capacidade de crescimento, nutri\ue7\ue3o e produ\ue7\ue3o de fen\uf3is totais inoculadas com fungos micorr\uedzicos arbusculares (FMAs). Realizou-se experimento em casa de vegeta\ue7\ue3o com tr\ueas tratamentos compostos por Gigaspora margarita , Glomus clarum e Glomus etunicatum testados de forma isolada e quatro tratamentos compostos pela combina\ue7\ue3o destas esp\ue9cies: Gigaspora margarita + Glomus clarum, Gigaspora margarita + Glomus etunicatum, Glomus clarum + Glomus etunicatum e Gigaspora margarita + Glomus clarum + Glomus etunicatum. Todos os tratamentos com o in\uf3culo f\ufangico foram avaliados sem a adi\ue7\ue3o de f\uf3sforo no substrato. Em compara\ue7\ue3o, foram utilizadas tr\ueas testemunhas (sem fungo) contendo tr\ueas doses de f\uf3sforo: 0, 50 e 100 mg dm-3, totalizando dez tratamentos. O delineamento experimental foi em blocos casualizados, com cinco repeti\ue7\uf5es. Ap\uf3s 140 dias de germina\ue7\ue3o, foi observado que todas as esp\ue9cies de FMAs inoculadas de forma isolada ou combinada promoveram benef\uedcios significativos no crescimento, nutri\ue7\ue3o e produ\ue7\ue3o de fen\uf3is totais nas mudas de cedro-australiano em solos contendo baixa disponibilidade de f\uf3sforo. A mistura no solo de Gigaspora margarita, Glomus clarum e Glomus etunicatum foi capaz de proporcionar incrementos significativos no desenvolvimento das plantas na maioria dos par\ue2metros avaliados, obtendose desempenhos superiores ou equivalentes \ue0s plantas testemunhas n\ue3o micorrizadas e cultivadas em solos adubados com f\uf3sforo. Isto sugere que os FMAs podem promover redu\ue7\ue3o na aduba\ue7\ue3o fosfatada no processo de produ\ue7\ue3o de mudas de cedro-australiano.This study aimed to evaluate the Australian red cedar seedlings for their ability to growth, nutrition and production of phenolic compounds under arbuscular mycorrhizal fungi (AMF) inoculation. An experiment was carried out under greenhouse conditions, with three treatments consisting of Gigaspora margarita , Glomus etunicatum , Glomus clarum in single inoculation, and four treatments composed by the combination of these species, Gigaspora margarita + Glomus clarum, Gigaspora margarita + Glomus etunicatum, Glomus clarum + Glomus etunicatum and Gigaspora margarita + Glomus etunicatum + Glomus clarum. All treatments with the fungal inoculum were evaluated without addition of phosphorus in the substrate. In comparison, three treatment controls were used (without fungus) containing three levels of phosphorus: 0, 50 and 100 mg dm-3, performing ten treatments. The experimental design was in random blocks with five replicates. After 140 days of germination, it was observed that all species of AMF inoculated alone or in combination, led to significant benefits on growth, nutrition and production of phenolic compounds in Australian red cedar seedlings in soils containing low phosphorus availability. The mixture in the soil of Gigaspora margarita, Glomus etunicatum and Glomus clarum was able to provide significant increases in plant growth in most parameters assessed, resulting in performance equivalent or higher than non-mycorrhizal and control plants grown in soil fertilized with phosphorus. This suggests that AMF can promote reduction in phosphorus addition during the production of Australian red cedar seedlings

The molecular structure of the borate mineral inderite Mg(H4B3O7)(OH)-5H2O - a vibrational spectroscopic study.

We have undertaken a study of the mineral inderite Mg(H4B3O7)(OH)_5H2O a hydrated hydroxy borate mineral of magnesium using scanning electron microscopy, thermogravimetry and vibrational spectroscopic techniques. The structure consists of ?B3O3?OH?5_2_ soroborate groups and Mg(OH)2(H2O)4 octahedra interconnected into discrete molecules by the sharing of two OH groups. Thermogravimetry shows a mass loss of 47.2% at 137.5 _C, proving the mineral is thermally unstable. Raman bands at 954, 1047 and 1116 cm_1 are assigned to the trigonal symmetric stretching mode. The two bands at 880 and 916 cm_1 are attributed to the symmetric stretching mode of the tetrahedral boron. Both the Raman and infrared spectra of inderite show complexity. Raman bands are observed at 3052, 3233, 3330, 3392 attributed to water stretching vibrations and 3459 cm_1 with sharper bands at 3459, 3530 and 3562 cm_1 assigned to OH stretching vibrations. Vibrational spectroscopy is used to assess the molecular structure of inderite

DYNAMICS OF VEGETATION IN AN ATLANTIC FOREST FRAGMENT IN NORTHEASTERN BRAZIL

Para testar a hip\uf3tese que a fisionomia, estrutura e din\ue2mica de esp\ue9cies lenhosas s\ue3o influenciadas pelo gradiente espacial borda-interior, foram coletados dados desses par\ue2metros em 2004, 2007 e 2010 em um fragmento de Floresta Atl\ue2ntica. Foram amostradas para o Dossel (DS), plantas com circunfer\ueancia do caule a altura do peito (CAP) > 15 cm e do sub-bosque (SB) com circunfer\ueancia ao n\uedvel do solo entre > 3 cm e CAP < 15 cm em um gradiente borda-interior formado h\ue1 35 anos. Em 2004, foram avaliadas tr\ueas posi\ue7\uf5es no fragmento: borda, intermedi\ue1ria e interior (> 150 m do limite florestal), sendo analisados 1000 m\ub2 por local, divididos em parcelas de 10 m\ub2, para medir os indiv\uedduos de maior classe. Em cada parcela foi instalada uma subparcela de 5 m\ub2 para amostragem do sub-bosque. Foram recolhidos ramos de tr\ueas indiv\uedduos adultos das esp\ue9cies, para obten\ue7\ue3o da densidade da madeira. O n\ufamero de indiv\uedduos (NI), \ue1rea basal (AB), taxa de mortalidade (TM), taxa de recrutamento (TR), incremento peri\uf3dico anual em di\ue2metro (IPA), taxa de perda (P) e ganho (G) de \ue1rea basal e as taxas de rotatividade em n\ufamero de indiv\uedduos (TN) e em \ue1rea basal (TA) da classe do componente arb\uf3reo nos tr\ueas locais evidenciaram n\ue3o haver efeito de borda significativo. Na borda, o sub-bosque teve menores valores de NI e AB e maiores TM, que no interior do fragmento. Diferen\ue7as entre os estratos acontecem, independentemente do selamento da borda, j\ue1 que plantas jovens na maior classe amostral apresentam indiv\uedduos no SB, que s\ue3o mais sens\uedveis que as \ue1rvores adultas \ue0s mudan\ue7as causadas pela cria\ue7\ue3o da borda. Taxas de perda e ganho de AB e de rotatividade em NI e AB, no SB, apresentaram valores maiores que no DS, o que aponta para um estrato com maiores mudan\ue7as. No DS n\ue3o ocorreram diferen\ue7as de riqueza de esp\ue9cies entre os ambientes, distinto do observado no SB. A composi\ue7\ue3o flor\uedstica no DS nos ambientes mais pr\uf3ximos ao limite florestal foi bastante distinta que a do interior, indicando que ainda n\ue3o houve recupera\ue7\ue3o da composi\ue7\ue3o flor\uedstica. Todos os ambientes do SB tiveram baixa similaridade. As bordas do DS e SB apresentaram maior concentra\ue7\ue3o de indiv\uedduos em intervalos de classe de menores valores de densidade de madeira. O DS j\ue1 recuperou a biomassa, embora os ambientes de borda e intermedi\ue1rio n\ue3o tenham recuperado a composi\ue7\ue3o de esp\ue9cies tardias. O SB ainda sofre efeito de borda. Possivelmente, a maior concentra\ue7\ue3o de indiv\uedduos em classe de menor densidade da madeira sendo resultado na borda.To test the hypothesis that the physiognomy, structure and dynamics of woody species are influenced by the spatial gradient edge-interior, data were collected from these parameters in 2004, 2007 and 2010 in an Atlantic forest fragment. Species were sampled for the Canopy (DS), with circumference > 15 cm to 1.3 m soil and understory (SB) in circumference between ground level> 3 cm and CAP <15 cm on edge-gradient formed within 35 years. In 2004, we assessed three positions in the fragment: edge, intermediate and inner (> 150 m from the forest boundary), and analyzed 1,000 m\ub2 per site, divided into plots of 10 m\ub2, to measure individuals of higher class. In each plot a sub-plot of 5 m\ub2 for sampling understory was installed. Branches were collected from three adult individuals from the species to obtain the density of the wood. The number of individuals (NI), basal area (BA), mortality (TM), recruitment rate (TR), periodic annual diameter increment (IPA), loss rate (P) and gain rate (G) Area and basal turnover rates in number of individuals (TN) and basal area (TA) class tree component in the three locations showed no significant edge effect. At the edge, the understory had lower values of NI and AB and higher TM than in the forest interior. Differences between the strata occur regardless of edge sealing, since young plants present in most individuals in the sample class SB, which are more sensitive than adult trees to changes caused by the creation of the edge. Rates of loss and gain of AB and AB turnover in NI and in SB presented higher values than on the DS, which points to a stratum with larger changes. In DS there were no differences in species richness among environments differs from that observed in SB. The floristic composition of the DS in environments closer to the forest limit was quite distinct from that of the interior, indicating that there has been no recovery of floristic composition. All environments SB had low similarity. The edges of the DS and SB had a higher concentration of individuals in class intervals smaller density values timber. The DS has already recovered biomass, although the edge and intermediate environments have not recovered species composition late. The SB still suffers the edge effect. Possibly, the highest concentration of individuals is in class with lower density wood result on edge

Antioxidant effects of oral Ang-(1-7) restore insulin pathway and RAS components ameliorating cardiometabolic disturbances in rats.

In prevention studies of metabolic syndrome (MetS), Ang-(1-7) has shown to improve the insulin signaling. We evaluated the HP?CD/Ang-(1-7) treatment on lipid metabolism, renin-angiotensin system (RAS) components, oxidative stress, and insulin pathway in the liver and gastrocnemius muscle and hepatic steatosis in rats with established MetS. After 7 weeks of high-fat (FAT) or control (CT) diets, rats were treated with cyclodextrin (HP?CD) or HP?CD/Ang-(1-7) in the last 6 weeks. FATHP?CD/ empty rats showed increased adiposity index and body mass, gene expression of ACE/ANG II/AT1R axis, and oxidative stress. These results were accompanied by imbalances in the insulin pathway, worsening of liver function, hyperglycemia, and dyslipidemia. Oral HP?CD/Ang-(1-7) treatment decreased ACE and AT1R, increased ACE2 gene expression. in the liver, and restored thiobarbituric acid reactive substances (TBARS), catalase (CAT), superoxide dismutase (SOD), insulin receptor substrate (Irs-1), glucose transporter type 4 (GLUT4), and serine/threonine kinase 2 (AKT-2) gene expression in the liver and gastrocnemius muscle improving hepatic function, cholesterol levels, and hyperglycemia in MetS rats. Overall, HP?CD/Ang-(1-7) treatment restored the RAS components, oxidative stress, and insulin signaling in the liver and gastrocnemius muscle contributing to the establishment of blood glucose and lipid homeostasis in MetS rats

Bioprospection of cellulolytic and lipolytic South Atlantic deep-sea bacteria

Background: Cellulases and lipases have broad industrial application, which calls for an urgent exploration of microorganisms from extreme environments as valuable source of commercial enzyme. In this context, the present work describes the bioprospection and identification of deep-sea bacteria that produce cellulases and lipases, as well their optimal temperature of activity. Results: The first step of this study was the screening of cellulolytic and lipolytic deep-sea bacteria from sediment and water column, which was conducted with substrates linked with 4-Methylumbelliferyl. Among the 161 strains evaluated, 40 were cellulolytic, 23 were lipolytic and 5 exhibited both activities. Cellulolytic and lipolytic bacteria are more common in sediment than at the water column. Based on the ability to produce cellulases and lipases three isolates were selected and identified (16S rRNA sequencing) as Bacillus stratosphericus , B. aerophilus and B. pumilus . Lipases of strain B. aerophilus LAMA 582 exhibited activity at a wide temperature range (4\uba to 37\ubaC) and include psychrophilic behaviour. Strain Bacillus stratosphericus LAMA 585 can growth in a rich (Luria Bertani) and minimal (Marine Minimal) medium, and does not need an inducer to produce its mesophilic cellulases and lipases. Conclusions: Deepsea sediments have great potential for bioprospection of cellulase and lipase-producing bacteria. The strains LAMA 582 and LAMA 585 with their special features, exhibit a great potential to application at many biotechnology process

Age-dependent effect of high-fructose and high-fat diets on lipid metabolism and lipid accumulation in liver and kidney of rats.

Background: The metabolic syndrome (MS) is characterized by variable coexistence of metabolic and pathophysiological alterations which are important risk factors for developing of type II diabetes and/or cardiovascular diseases. Increased of MS patients in worldwide has stimulated the development of experimental models. However, it is still challenging to find an dietetic model that most closely approximates human MS and, in addition, is not yet fully established the effect of different diets of MS in lipid metabolism in rats of different ages. The aim of this study was to evaluate the effect of different diets of MS in lipid metabolism and ectopic fat deposition and define the most appropriate diet for inducing the characteristic disturbances of the human MS in rats of different ages. Methods: Young (4 weeks old) and adult rats (12 weeks old) were given a high-fat (FAT) or high-fructose diet (FRU) for 13 weeks and biochemical, physiological, histological and biometric parameters were evaluated. Results: In young rats, the FAT diet induced increased mean blood pressure (MAP) and heart rate (HR), body weight after 6 to 10 weeks, and in the 13th week, increased the liver, mesenteric, retroperitoneal and epididymal fat weights,fasting glucose, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and reduced HDL cholesterol; and also induced non-alcoholic fatty liver disease (NAFLD) and renal inflammatory infiltrates. In adult rats, the FRU diet induced transient elevations of MAP and HR in the 6th week, and, at 13 weeks, increased fasting glucose, triglycerides, total cholesterol, AST and ALT; increased liver, kidneys and retroperitoneal fat weights; and induced macrovesicular and microvesicular NAFLD, the presence of fat cells in the kidney, glomerular sclerosis, and liver and kidney inflammation. Additionally, the FAT and FRU diets induced, respectively, increases in liver glycogen in adults and young rats. Conclusions: Our data show that FRU diet in adult rats causes biggest change on metabolism of serum lipids and lipid accumulation in liver and kidney, while the FAT diet in young rats induces elevation of MAP and HR and higher increased visceral lipid stores, constituting the best nutritional interventions to induce MS in rats

Bioconversion of cassava starch by-product into Bacillus and related bacteria polyhydroxyalkanoates

Background: Unlike petroleum-based synthetic plastics, biodegradable biopolymer generation from industrial residue is a key strategy to reduce costs in the production process, as well as in the waste management, since efficient industrial wastewater treatment could be costly. In this context, the present work describes the prospection and use of bacterial strains capable to bioconvert cassava starch by-product into biodegradable polyhydroxyalkanoates (PHAs). Results: The first step of this study was the bacterial competence screening which was conducted with 72 strains covering 21 Bacillus and related species. The microorganism growth in a medium with a starch substrate was measured by an innovative MTT assay, while the ability of the bacteria to secrete amylase and produce PHA was evaluated by the Nile Red Dye method. Based on growth and potential for PHA production, four isolates were selected and identified as Bacillus megaterium by 16S rRNA sequencing. When cultivated in hydrolyzed cassava starch by-product, maximum production reached 4.97 g dry biomass/L with 29.7% of Poly-(3-hydroxybutyrate) (characterized by FTIR). Conclusions: MTT assay proved to be a reliable methodology for monitoring bacterial growth in insoluble media. Selected amylolytic strains could be used as an alternative industrial process for biodegradable plastics production from starchy residues, reducing costs for biodegradable biopolymer production and wastewater treatment operations